We report the development of a molecular probe for in vivo high-resolution imaging of temperature concurrently with oxygen. The probe is based on a chromophore capable of microsecond-scale emission occurring simultaneously from two well spectrally resolved excited electronic states, existing in thermal equilibrium with one-another: phosphorescence and delayed fluorescence. While the emission decay time is sensitive to both temperature and oxygen, the ratio of the decay intensities is sensitive only to temperature. Because ratiometric measurements (i.e. the ratio of decay intensities at two different wavelengths) in vivo are jeopardized by optical heterogeneities of the medium (differential absorption and scattering by endogenous tissue chromophores), the probe is supplemented by a second molecule of nearly identical structure, but emitting regular prompt fluorescence at the wavelengths matching those of the phosphorescence and delayed fluorescence of the probe. The second chromophore is excited simultaneously with the probe, serving as a fluorescent correction standard to account for absorption/scattering uncertainty, since prompt fluorescence is independent of temperature and/or oxygen. The microsecond-scale emission of the probe and the nanosecond emission of the standard are measured simultaneously, taking advantage of their significantly different time scales. Synthesis, photophysical characterization and in vivo tests will be discussed.

Phosphorescence lifetime-based probe for oxygen and temperature

Macromolecular Chemistry: The Second Century

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ACS Spring 2020 National Meeting & Expo

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